The TurboCapture 96 mRNA Kit uses unique oligonucleotide immobilization technology to provide a fast and simple procedure for purifying mRNA. Cell lysates are added to the wells of a TurboCapture plate (96 wells), and mRNA is allowed to hybridize to the immobilized oligo-dT in each well. Contaminants are washed away, and the isolated mRNA is then either used directly in cDNA synthesis or eluted for use in downstream applications. For higher throughput purification in 384-well format, the TurboCapture 384 mRNA Kit is available.

The TurboCapture 96 mRNA Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Performance

TurboCapture mRNA Kits provide rapid and easy purification of mRNA from cultured cells, including adherent and suspension cells (see figure "mRNA purification from suspension cells"). Due to the standard plate formats and simple workflows, TurboCapture mRNA Kits can be automated, allowing mRNA purification, cDNA synthesis, and PCR to be performed in the same well. This avoids the need for sample transfer and ensures high well-to-well reproducibility (see figure "High well-to-well reproducibility"). When immobilized oligo-dT in the well is used as primer, the synthsisized cDNA is covalently linked to the well and can be reused several times (see figure "Sequential PCR analysis"). The kits are ideally suited for high-throughput gene expression analysis applications, such as screening of siRNAs by real-time RT-PCR (see figure "Parallel analysis of siRNAs"). TurboCapture mRNA Kits also provide high sensitivity in gene expression analysis, allowing detection of targets from as little as a single cell (see figure "Gene expression analysis").

Principle

Rapid, cost-effective mRNA purification from adherent and suspension cells, as well as from total RNA, is achieved with minimal hands-on time using the unique oligonucleotide immobilization technology in TurboCapture mRNA Kits. The kits allow mRNA to be purified by hybridizing to immobilized oligo-dT in each well of a 96- or 384-well plate. The immobilized oligo-dT in the well can be used as primer, and cDNA is synthesized while mRNA remains hybridized, yielding cDNA which is covalently linked to the well. TurboCapture plates are compatible with most thermal cyclers, allowing mRNA hybridization, and subsequent cDNA synthesis and PCR, to take place in the same plate, minimizing sample handling and reducing consumable costs. The procedure can be automated on robotic instruments, due to the standard plate formats and simple workflows.

Procedure

With the TurboCapture mRNA Kits, mRNA can be purified from total RNA, as well as from a range of cell types, including adherent and suspension cells. (If purifying mRNA from suspension cells, Buffer TCL, 2x [cat. no. 1031586] must also be purchased.) Simply add sample lysates to the wells of a TurboCapture plate (96 or 384 wells). Allow mRNA to hybridize to the immobilized oligo-dT in each well, and wash away the contaminants. The isolated mRNA can be used directly in cDNA synthesis or, alternatively, can be eluted for use in downstream applications (see flowchart "TurboCapture mRNA procedure"). The immobilized oligo-dT in the well can be used as a primer for cDNA that will be covalently linked to the well and can be reused. Alternatively, soluble oligo-dT primers or random hexamers can be used to prepare soluble cDNA (see figure "mRNA purification and cDNA synthesis in the same well").